1.Impacts of cordycepin on isoproterenol-induced cardiomyocyte apoptosis by regulating cAMP/PKA signaling pathway
Juan LI ; Zonghan ZHAO ; Chunhou HU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(12):1489-1494
Objective To investigate the impacts of cordycepin(Cor)on isoproterenol(ISO-induced cardiomyocyte apoptosis and cyclic AMP/protein kinase A(cAMP/PKA)signaling path-way.Methods After H9c2 cardiomyocytes were treated with ISO(10 μmol/L),the cells were further treated with Cor(1,5,10,15 μg/ml),respectively to detect cell proliferation.Then the H9c2 cells were grouped into Control group,ISO group,5 μg/ml Cor group(Cor-L group),10μg/ml Cor group(Cor-H group),and 10 μg/ml Cor+20 μmol/L H-89 group(combined group).Flow cytometry was applied to detect cell apoptotic rate in above groups.Phalloidin staining was used to detect the surface area of the cells.RT-PCR was used to detect the expression of hypertro-phy related genes,such as ANP,BNP,and β-myosin heavy chain(β-MHC)in the cardiomyocytes.ELISA was conducted on test the levels of inflammatory factors,TNF-α,IL-6 and IL-1β and cAMP.Western blot assay was employed to measure the expression levels of B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),Caspase-3,and PKA.Results Compared with the Control group,the apoptotic rate[(33.89±3.68)%vs(2.57±0.26)%],cell surface area,and the expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC were in-creased,and the expression levels of Bcl-2,cAMP,and PKA were decreased in the ISO group(P<0.05).The Cor-L and Cor-H groups had significantly lower apoptotic rate[(20.58±1.93)%and(7.58±0.76)%vs(33.89±3.68)%],smaller cell surface area,and decreased expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC,and increased levels of Bcl-2,cAMP,and PKA when compared with the ISO group(P<0.05).The apoptotic rate[(28.69±2.82)%vs(7.58±0.76)%],cell surface area,and expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC were elevated,and the expression levels of Bcl-2,cAMP and PKA protein were decreased in the combined group than the Cor-H group(P<0.05).Conclusion Cor inhibits ISO-induced cardiomyocyte apoptosis by activating the cAMP/PKA signaling pathway.
2.Impacts of cordycepin on isoproterenol-induced cardiomyocyte apoptosis by regulating cAMP/PKA signaling pathway
Juan LI ; Zonghan ZHAO ; Chunhou HU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(12):1489-1494
Objective To investigate the impacts of cordycepin(Cor)on isoproterenol(ISO-induced cardiomyocyte apoptosis and cyclic AMP/protein kinase A(cAMP/PKA)signaling path-way.Methods After H9c2 cardiomyocytes were treated with ISO(10 μmol/L),the cells were further treated with Cor(1,5,10,15 μg/ml),respectively to detect cell proliferation.Then the H9c2 cells were grouped into Control group,ISO group,5 μg/ml Cor group(Cor-L group),10μg/ml Cor group(Cor-H group),and 10 μg/ml Cor+20 μmol/L H-89 group(combined group).Flow cytometry was applied to detect cell apoptotic rate in above groups.Phalloidin staining was used to detect the surface area of the cells.RT-PCR was used to detect the expression of hypertro-phy related genes,such as ANP,BNP,and β-myosin heavy chain(β-MHC)in the cardiomyocytes.ELISA was conducted on test the levels of inflammatory factors,TNF-α,IL-6 and IL-1β and cAMP.Western blot assay was employed to measure the expression levels of B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),Caspase-3,and PKA.Results Compared with the Control group,the apoptotic rate[(33.89±3.68)%vs(2.57±0.26)%],cell surface area,and the expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC were in-creased,and the expression levels of Bcl-2,cAMP,and PKA were decreased in the ISO group(P<0.05).The Cor-L and Cor-H groups had significantly lower apoptotic rate[(20.58±1.93)%and(7.58±0.76)%vs(33.89±3.68)%],smaller cell surface area,and decreased expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC,and increased levels of Bcl-2,cAMP,and PKA when compared with the ISO group(P<0.05).The apoptotic rate[(28.69±2.82)%vs(7.58±0.76)%],cell surface area,and expression levels of TNF-α,IL-6,IL-1β,Bax,Caspase-3,ANP,BNP,and β-MHC were elevated,and the expression levels of Bcl-2,cAMP and PKA protein were decreased in the combined group than the Cor-H group(P<0.05).Conclusion Cor inhibits ISO-induced cardiomyocyte apoptosis by activating the cAMP/PKA signaling pathway.

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